JP2005508893A - Use of fucans in the treatment of adhesions, arthritis and psoriasis - Google Patents

Abstract

To treat adhesions, arthritis and psoriasis more effectively with few side effects.
Fucans are administered in various forms.

Description

Cross-reference to related applications

  This application claims priority from US Provisional Patent Application No. 60 / 315,362, filed Aug. 29, 2001.

  Adhesions resulting from surgery (surgical adhesions) are a type of scar that usually forms between two parts of the body after surgery. Adhesions can cause serious problems. For example, adhesions on female reproductive organs (ovary, fallopian tube) can cause infertility, sexual pain (painful combination), and severe pelvic pain. Adhesions that occur in the intestine can cause intestinal obstruction or bowel obstruction. Adhesions can also occur around the heart, around the spine, and other sites such as the hands. In addition to resulting from surgery, adhesions can also be formed by events such as endometriosis, infection, chemotherapy, radiation and cancer.

  Adhesion and other angiogenesis-related diseases such as arthritis and psoriasis can last for weeks, months or years, and the treatment is long-term and expensive. See Robbins Pathological Basis of Disease by Cotran, R.S., Kumar, V., Robbins, S.L., p75 (W. B. Saunders Co., 1989). Such diseases and symptoms can develop into chronic inflammatory symptoms with severe consequences for both the mental and physical health of the patient. To make matters worse, there are few treatment options for patients suffering from surgical adhesions, arthritis and psoriasis. Often patients are treated with drugs such as steroidal or non-steroidal anti-inflammatory drugs to alleviate the symptoms of the disease. However, such treatments are unlikely to have an appropriate effect over a long period of time, and conversely, if used frequently, serious side effects (such as gastric ulcers caused by non-steroidal anti-inflammatory drugs, or steroids) More serious toxicity due to the abuse of Other, more potent anti-proliferative and / or anti-angiogenic agents such as paclitaxel, methotrexate, doxorubicin, and etoposide will show potent therapeutic modalities, but are not life-threatening diseases These powerful drugs against the disease can only be used to a limited extent due to undesired toxicity and side effects.

  However, there remains an unmet need for compounds, compositions, methods, etc. (including delivery methods) to more effectively treat one or more of these diseases, preferably with few side effects. Therefore, it is an object of the present invention to provide compounds, compositions, methods, etc. that realize one or more such advantages.

  In order to solve the above problems, the present invention provides compositions and methods comprising fucans such as fucoidan for the treatment of surgical adhesions, arthritis and psoriasis. Fucans show important therapeutic effects for any of these diseases, but have few side effects.

  In one aspect, the present invention provides a method of treating adhesions in an animal, which can be a human or other desired subject. The method of treatment involves administering a therapeutically effective amount of fucan (which may be fucoidan) to a disease site that may have adhesions. The disease site may be a surgical site and the fucan may be delivered directly to the disease site as a composition. Fucan may be administered in a sustained manner from a polymeric dosage form in a sustained release, substantially continuously to the disease site. The polymer dosage form may be a film, patch, paste, microsphere, implant, gel, spray or liquid. Fucans may be administered as a pharmaceutical composition comprising a form comprising at least one of creams, pastes, injectable excipients and polymers. (Except where otherwise specified or apparent from the context, all of the “embodiments”, “side surfaces”, “features”, and the like can be combined or replaced in any manner.)

  The fucan may be administered as a pharmaceutical composition comprising fucan and at least one therapeutically effective amount of another agent. Other drugs include paclitaxel, doxorubicin, camptothecin, etoposide, mitoxantrone, methotrexate, menadione, plumbazine, juglone, β-lapachone cyclosporine, sulfasalazine, steroids, rapamycin, retinoids, docetaxel and colchicine, antisense oligonucleotides, ribozymes And at least one of them. A therapeutically effective amount of fucan may be delivered as part of the composition, with fucan ranging from approximately 0.1% to approximately 35%, approximately 5% to approximately 50%, approximately 20% to approximately 80%, approximately 80%. % To about 100% w / v may be included in the composition.

  The composition may further comprise at least one pharmaceutically acceptable excipient such as pluronic, cellulose, alginate, acrylate, hyaluronic acid, polyethylene glycol, injectable excipient and chitosan and the like. Fucan is orally administered, administered directly to the diseased site, administered by injection to the diseased site, intraocular administration, intraperitoneal administration, intramuscular administration, intraarticular administration, intralesional administration, subcutaneous administration, intravaginal administration, rectal administration Or it can administer by local administration and other arbitrary methods.

  In other aspects, the methods of the invention comprise treating arthritis, psoriasis, or angiogenic eye disease, and administering a therapeutically effective amount of fucan to the disease site.

  In a further aspect, the invention provides a therapeutically effective amount of fucan and at least one pharmaceutically selected from the group consisting of pluronic, alginate, acrylate, hyaluronic acid, polyethylene glycol, injectable excipients and chitosan. Pharmaceutical compositions comprising a fucan polymeric dosage form comprising an acceptable excipient are provided. The polymer dosage form may be composed of a film, paste, microsphere, spray, lotion, liquid or implant or other desired form. The pharmaceutical composition may further comprise at least one therapeutically effective amount of other agents such as antisense oligonucleotides, ribozymes and oligonucleotide RNA inhibitors.

  The composition can be used in the manufacture of a medicament for treating adhesions such as surgical adhesions, arthritis, psoriasis or other desired diseases. The present invention further provides a method for producing a medicament capable of alleviating a symptom associated with at least one of adhesions, arthritis and psoriasis in a human patient. This method of production involves mixing a therapeutically effective amount of fucan (such as fucoidan) and a pharmaceutically acceptable excipient or buffer.

  These and other aspects, features, and embodiments are disclosed in this application, including the following detailed description and the accompanying drawings. Furthermore, this document lists various references (including cross-references to related applications) discussing specific systems, devices, methods, and other information. All such references are hereby incorporated by reference in their entirety, ie, all of their teachings and disclosures, whether or not explicitly stated in this document.

  The present invention includes compositions and methods including approaches to inhibit cell proliferation, inflammatory response and angiogenesis using sulfated polysaccharides known as fucans, including surgical adhesions (adhesions resulting from surgery), Can be used to treat arthritis and psoriasis. Fucans such as fucoidan appear to inhibit neutrophil activation, inhibit release of inflammatory enzymes from arthritis-related cells, and inhibit angiogenesis in chick membranes and surgical adhesions. Since all cells have a fucose-binding receptor, in some embodiments, the fucan is at the site of the disease in order to substantially continuously expose the target tissue with a fucan (such as fucoidan) in a sustained release manner from the polymeric dosage form. Delivered directly to. Fucan can have multiple effects in vivo (especially affecting thrombin and complement in the blood), so site-specific sustained release of fucan is an alternative to systemic administration that can reduce hematologic toxicity It becomes a means.

  In the following, first a general description of fucans, adhesions, arthritis and psoriasis will be given, then several embodiments of the invention will be described, and finally several examples will be described.

  General background on fucans, adhesions, arthritis and psoriasis

  Fucans

  Fucans (including fucoidan) are high molecular weight sulfate polysaccharides extracted from brown algae. This compound has been reported to have multiple inhibitory activities in vivo and in vitro, including antithrombin, antiproliferation, anticomplement, anticancer and antineutrophil migration effects. Fucans may contain cell-cell junctions via integrin-selectin molecules or may block various binding events that occur on the cell surface by binding to thrombin or complement in the blood or fucose receptors on the cell surface. unknown.

  Such activity is believed to contribute to anti-inflammatory properties based on inhibiting (for example) the binding of lymphocytes or neutrophils to vascular endothelial cells, which infiltrate the tissue compartment. It seems to prevent the subsequent inflammation. See Patankar, M. S., et al., J. Biol. Chem. 268: 21770-21776 (1993); Brandley, B. K., et al., J. Cell Bio1. 105: 991-997 (1987). Recent studies have also shown that fucans inhibit vascular smooth muscle cell proliferation. See Logeart, D., et al., Eur. J. Cell Biol. 74: 376-384 and 385-390 (1997). This suggests that fucans may have anti-restenosis ability (but not demonstrated). Fucans have been shown to be slowly taken up into cells after surface binding to endothelial and smooth muscle cells. See Glabe, C. G., et al., J. Cell Science 61: 475-490 (1983); Logeart, D., et al., Eur. J. Cell Biol. 74: 376-384 (1997).

  Riou, D., et al., Anticancer Res., 16 (3A): 1213-1218 (1996); Itoh, H., Anticancer Res., 13 (6A): 2045-2052 (1993); Nishiro, T. , et al., Thromb. Res., 62: 765-773 (1991); Blondin, C., et al., Mol. Immunol., 31: 247-253 (1994); Patankar, MS, et al., J. Biol. Chem., 268: 21770-21776 (1993). In Japan, fucoidan extracted from various seaweeds is sold as a health food. Fucoidan has been proposed as a cosmetic or dermatological agent. See JP 01031707 and JP 01085905. Fucoidan has also been reported as a potential anticancer agent. See Riou. D., Anticancer Res. 16: 3a 1213-18 (1996); Itoh, H., et al., Anticancer Res., 15: 5b 1937-47 (1995). Fucoidan has been reported not to inhibit angiogenesis in vitro. See Soeda, S., et al., Biochim. Biophysica Acta (1): 127-134 (2000). Similarly, fucoidan was also found to stimulate serum-induced (in vitro) HUVE cell proliferation. This suggests the possibility of having a pro-angiogenic action (effect) (even though inhibition may have occurred in the presence of fibroblast growth factor). See Giraux, J., et al., Eur. J. Cell Biol. 77 4: 352-9 (1998). Several studies have shown that fucans inhibit endothelial cell monolayer binding. See Glabe, C. G., J. Cell Science, 61: 475-490 (1983). Since the cells that make up the capillaries are endothelial cells, this report may inhibit certain aspects (stages) of cell adhesion in vitro, but these data indicate that the anti-angiogenic effects of fucoidan in vivo It shows that it has not been proven at all. Fucoidan has been reported to inhibit the binding of Helicobacter to gastric cells. This suggests that it has an anti-gastric ulcer action. See Shibat, H. J., Nutr. Sci. Vitaminol. 45: 325-336 (1999).

  There are also reports on other sulfated polysaccharides (including branched and straight chain) with specific anticoagulant activity. See Pereira, M. S., J. Biol. Chem. 12: 7656-67 (1999). Dextran sulfate and its derivatives inhibit cancer cell growth (see Bittoun, P., Carbohydrate Res. (3-4): 247-255 (1999)) and have anticoagulant activity (Mauray, S ., J. Biomat. Sci. Poly ed. 9: 373-87 (1998)) was also reported. Sulfuric acid polysaccharides have also been proposed as antiviral agents for use, for example, against AIDS. See EP 00293826; JP 01313433.

  adhesion

  The formation of adhesions is a complex process in which tissues that are normally separated in the body proliferate and enter each other. Surgical adhesions (also known as post-surgical adhesions) result from normal wound healing responses elsewhere in the tissue to trauma, and in abdominal surgery occur in more than two-thirds of the entire patient. See Ellis, H., Surg. Gynecol. Obstet. 133: 497 (1971); Wiebel, M-A. And Majno, G., Am. J. Surg. 126: 345 (1973). The outcome of such adhesions will vary depending on the relevant surgical site. Problems such as pain, infertility, bowel obstruction can occur, and even the risk of death after cardiac surgery can be increased. diZerega, GS, Prog. Clin. Biol. Res. 381: 1-18 (1993); diZerega, GS, Fertil. Steril. 61: 219-235 (1994); Dobell, AR, Jain, AK, Ann. Thorac. See Surg. 37: 273-278 (1984).

  The adhesion formation process initially involves the formation of a fibrin skeleton and normal tissue repair. In normal repair processes, fibrinolysis is expected along with mesothelial repair. However, in surgical adhesion formation, as the fibrin matrix matures, fibroblasts grow into the network and angiogenesis occurs. As a result, organized adhesions are established within 3-5 days. See Buckman, R.F., et al., J. Surg. Res. 21: 67-76 (1976); Raferty, A. T., J. Anat. 129: 659-664 (1979).

  Inflammatory processes include neutrophil activation in trauma tissue, fibrin deposition and connection between adjacent tissues, macrophage invasion, fibroblast proliferation to the area, collagen deposition, angiogenesis, establishment of permanent adhesion tissue. At present, prophylactic treatment includes prevention of fibrin deposition, alleviation of inflammation (steroidal and non-steroidal anti-inflammatory drugs) and removal of fibrin deposits.

Interventional attempts to prevent the formation of post-surgical adhesions included the use of water flotation or barrier devices. The water suspension method is described in dextran (see Adhesion study group, Fertil. Steril. 40: 612-619 (1983)) or carboxymethylcellulose (Elkins, TE, et al., Fertil. Steril. 41: 926-928 (1984)). An attempt is made to keep each organ or organ away from each other by dripping a large amount of polymer solution such as Complete resorption from oxidized regenerated cellulose (Interceed ^™ ), synthetic barrier membrane made from polytetrafluoroethylene (Gore-tex surgical membrane) and modified hyaluronic acid / carboxymethylcellulose (HA / CMC) mixture (Seprafilm ^™ ) Sexual membranes have also been used to reduce postoperative adhesion formation in animals and humans. Burns, JW, et al., Eur. J. Surg. Suppl. 577: 40-48 (1997); Burns, JW, et al., Fertil. Steril. 66: 814-821 (1996); Becker, JM, et al., J. Am. Coll. Surg. 183: 297-306 (1996). The success of this HA / CMC membrane may be due to its ability to separate tissue during the peritoneal wound repair process as adhesions are formed. This film was observed to form a transparent viscous film on the damaged tissue for 3-5 days after application. This period of 3 to 5 days corresponds to a period corresponding to the time course of postoperative adhesion formation. See Ellis, H., Br. J. Surg. 50: 10-16 (1963). Intraperitoneal administration of anti-inflammatory agents such as dexamethasone or corticosteroids hardly inhibited adhesion formation. See diZerega, GS, Fertil. Steril. 61: 219-235 (1994); Hockel, M., Ann. Chir. Gynecol. 76: 306-313 (1987).

  arthritis

  Arthritis, such as rheumatoid arthritis (RA), is a debilitating chronic inflammatory disease affecting approximately 2% of the world's population. The symptoms are characterized by pain, swelling, synoviocyte proliferation (pannus formation), angiogenesis and joint tissue destruction. As the disease progresses, arthritis often damages vital organs and can even lead to death. Arthritis involves many components of the immune system (macrophages / monocytes, neutrophils, B cells and T cells), complex cytokine responses and synovial cell dysfunction and proliferation. Early intensive treatment with a combination of slow-acting anti-rheumatic drugs (DMARDS) such as methotrexate and cyclosporine or azathioprine is now recommended. See Arthritis and Rheumatism, 39 (5): 713-722 (1996).

  Crystal-induced arthritis affects approximately 1% of the population and is characterized by crystal-induced activation of macrophages and neutrophils in the joints, and unbearable pain continues for days. As crystal-induced arthritis progresses, the interval between onsets becomes shorter and the patient's condition reaches an unacceptable level. Crystal-induced arthritis is generally treated with an NSAID depending on the symptoms. For a more detailed description of the pathophysiology of crystal-induced arthritis and other forms of inflammatory arthritis, see McCarty, et al., Arthritis and Allied Conditions by Lea and Febiger, Philadelphia 1495 (1985).

  psoriasis

  Psoriasis is a common chronic inflammatory skin disease characterized by raised, hypertrophic and scaly lesions that cause itching, tingling, stinging and easy bleeding. More than 2% of Americans suffer from psoriasis, and patients often have concomitant arthritic symptoms. The cause of psoriasis is unknown, and there is currently no cure for psoriasis. There is also evidence to support the concept of autoimmune disease. Psoriasis is further characterized by neutrophil activation, cell proliferation and angiogenesis.

  Skin cells are thought to follow two growth pathways: normal growth and wound healing. In normal growth, cells develop in the basal layer and migrate through the epidermis to the skin surface. Dead cells fall off the skin surface as fast as new cells form below them. During wound healing, acceleration of proliferation and repair is induced, resulting in rapid turnover of skin cells, increased blood supply and inflammation. In a sense, psoriasis is an excessive wound healing process. If the skin does not shed the skin cells (keratinocytes) as fast as the skin cells are made, a bulge (accumulation) will occur. As such, scaly lesions and angiogenesis (and increased blood supply) will occur. At the same time, lymphocytes, neutrophils and macrophages will cause tension, swelling and inflammation. Current drug treatments include the use of steroidal and non-steroidal anti-inflammatory agents to treat inflammatory conditions. Methotrexate and cyclosporine are also used with only a minor effect. The current cost of treating psoriasis in the United States has reached more than $ 3 billion annually.

  Outline

  The present invention provides fucans (including derivatives and analogs thereof) for the treatment or prevention of surgical adhesions (adhesions resulting from surgery), rheumatoid arthritis and psoriasis (hereinafter referred to as “treatment”). Includes treatment of symptoms that are currently onset and prevention of later onset of symptoms). As shown in the examples below, fucans (and especially fucoidan) inhibit cell proliferation, inflammatory responses / phenomena and angiogenesis, such as appearing in surgical adhesions.

  In one embodiment, fucans such as fucoidan are used to inhibit or prevent angiogenesis. In other embodiments, fucans such as fucoidan inhibit or prevent activation of inflammatory cells such that cells that cause an inflammatory response at the disease site are suppressed. This is important because many diseases, such as osteoarthritis and the like, are not necessarily associated with the accumulation of inflammatory cells at the disease site. Therefore, the use of such fucans can inhibit or prevent longer-lasting activation of resident macrophages, neutrophils and other inflammation-inducing cells that cause the undesirable chronic effects of the disease. it can. Such fucan activity applies in the present invention to surgical adhesions, arthritis and psoriasis.

  In one embodiment of the invention, the fucans (including derivatives and analogs thereof) are configured as sustained release dosage forms so that the effective concentration of the drug to be delivered to the disease site can be maintained. Can do. In other embodiments, fucoidan is used to treat surgical adhesions. A specific experiment was conducted on fucoidan. Such examples show the inhibitory effect of fucoidan on primitive (primary) chondrocytes derived from fresh cartilage (cells involved in rheumatoid arthritis). This suggests that this agent has potential as an anti-arthritic agent. In particular, the clear ability of fucoidan to inhibit collagenase and stromelysin production provides a cure for cases where the release of these and / or other metalloproteinases causes medical problems.

  In other embodiments, fucoidan can be used in combination with other therapeutic agents to anticipate good efficacy against less toxic disease processes. For example, in the treatment of surgical adhesions, powerful anti-proliferative agents such as doxorubicin, camptothecin, etoposide, mitoxantrone, methotrexate, menadione, plumbazine, juglone, β-lapachone cyclosporine, sulfasalazine, steroids, rapamycin, retinoids, paclitaxel, Docetaxel, colchicine, and other microtubule inhibitors, and other analogs and derivatives thereof, may have undesired toxicity at drug concentrations required to inhibit the adhesion process in the absence of fucan, but fucoidan, etc. In combination with these fucans, efficacy can be achieved at lower concentrations to achieve the desired result.

  In another embodiment, the fucan itself may be a dosage form (formulation form). For example, fucans should be made in the form of a thin film that can be applied to the tissue (affected area) continuously by applying an effective concentration of the agent to the tissue (affected area) by applying it directly to the surgical trauma site and slowly dissolving (exuding) the fucan. Can do. In fact, such a dosage form (dosage form) is a sustained release drug delivery for itself (as an active ingredient) or for other agents (such as paclitaxel) that can be incorporated into the dosage form. Can act as a system. Fucans can also be configured in the form of tablets, capsules, microspheres (microspheres), pastes, gels, powders, and fumes, and can be administered orally or rectally as solids or liquids. it can.

  In general, fucans are applied alone or in compositions by applying or injecting as pastes, gels, sprays, (fine) particles, films, solutions (aqueous solutions), liquids (liquids), lotions, creams or implants. Can be administered as part. For the administration route / site, oral administration, systemic administration, intraocular administration, subcutaneous administration, intraperitoneal administration, intramuscular administration, intraarticular administration, intralesional administration, intravaginal administration, intrarectal administration or (for example, patch (patch) )) Topical administration. These routes of administration may be the site of action of the dosage form of the fucan or fucan-drug composition in certain cases. A therapeutically effective amount of fucan can also be delivered as part of the composition and is about 5% to about 50%, about 20% to about 80%, about 80% to about 100% w / v ( Ratio). Fucans may be placed in a suitable container or container, then in a kit and further labeled, preferably a label indicating the approval of a government regulatory authority (eg, the Food and Drug Administration of the United States of America).

  To treat adhesions, fucans or fucan-containing compositions can be used in affected areas or surgery in the form of solutions, (fine) particles, suspensions, films, pastes, gels, sprays, liquids, lotions, implants or other desired forms. Can be administered directly to the site. Adhesion can also be treated by systemic delivery of fucans using intravenous, subcutaneous, intramuscular, intraperitoneal, oral, or other desired routes of administration. To treat arthritis, fucans or fucan-containing compositions can also be injected directly into the joints in pastes, gels, sprays, liquids, lotions, solutions, suspensions or other desired forms. Treatment or prevention of arthritis can also be performed by systemic delivery of fucan following the routes of intravenous administration, intramuscular administration, intraperitoneal administration, subcutaneous administration or oral administration.

  In one aspect, the invention also applies to the treatment of ocular neovascular diseases. For example, diabetic retinopathy is a complication of diabetes that can cause blindness such as blurred vision or destruction of the retina caused by damage to the blood vessels of the retina and the growth of new blood vessels (angiogenesis). . Macular degeneration is caused by the invasion of new blood vessels under the retina and is the leading cause of blindness in the United States and Europe, where only 200,000 new cases occur annually in the United States and only 15% can be treated with current laser therapy. is there. The present invention, in certain embodiments, provides a pharmacological approach for treating these diseases using the methods and compositions described herein adapted for ocular application. Such system modifications include dosage forms or pluronic, alginate, acrylate, cellulose, hyaluronic acid, polyethylene glycol, chitosan, and analogs or derivatives thereof, and various other pharmaceutically acceptable molding agents. Includes chemical cross-linking that slows the dissolution (leaching) rate from the mixture with other excipients.

  In yet other embodiments, the fucans constitute a charged aqueous gel with a positively charged excipient such as chitosan or poly-1-lysine. For example, agents such as antisense oligonucleotides, ribozymes and oligonucleotide RNA inhibitors can be incorporated into such gels for administration to disease sites. Alternatively, such a drug-containing gel or a drug dissolved in a fucan solution can be dried and pulverized into particles (fine particles). This particle can be administered to a diseased site and function as a sustained release dosage form, or the particle can function as a transfer agent because fucans bound to the surface are taken into cells. it can. The administration of such particles is such as pluronic, cellulose, alginate, acrylate, hyaluronic acid, polyethylene glycol, chitosan, injectable excipients, and analogs or derivatives thereof, and various other polymeric excipients. It can be further facilitated by using pharmaceutically acceptable excipients such as excipients.

  With regard to transfection and the combined use of fucans and nucleic acid sequencing agents, the advanced medical field known as gene therapy is limited by drug delivery problems, but ribozymes, antisense nucleotides and oligonucleotide RNA inhibitors Gene fragments or nucleic acid strands such as oligonucleotides containing can sometimes be inhibited from taking up into cells due to the charge and large molecular weight of these compounds. Recently, it has been proposed to use microparticles (eg calcium phosphate) containing genes or nucleic acids as transfection agents so that they bind to the cell surface and are taken up by food or drink or invaginated to introduce genes or nucleic acids into cells. . Many cells have fucose receptors on their membrane surfaces. The present invention provides the use of fucans as transfection agents for nucleic acid strands. In one embodiment, the nucleic acid strands can be bound or encapsulated within fucoidan microparticles that are dissolved (exuded) before being administered (arrived) to the target cell site. To prevent, it can be chemically cross-linked.

  Fucans can be used alone or in combination with other drugs in combination with body implants. Such implants include, but are not limited to, various medical devices such as catheters, shunts, membranes, stents, sponges, fillers, prosthetic joints and portions thereof, and orthopedic implants. Such implants (implantation materials) can include or be coated with fucans alone or in combination with fucans and other drugs and excipients.

  Except for the claims, unless stated otherwise, “or (or)” includes “and (and)” and “and (and)”. The description also includes “or”. Non-limiting terms should not be construed as limiting unless explicitly stated or otherwise clearly indicated by context. (For example, “including,” “having,” and “comprising” typically includes “but is not limited to”. Singular forms such as “one (a)”, “one” (an) and “(the) said or the above” or “the”) are intended to be in this context. In some cases, this may mean a plural unless explicitly stated or otherwise clearly indicated by context.

  The scope of the system and method of the present invention includes the concepts “means plus function” and “step plus function”. However, these terms described in this application are the so-called “means plus function” in the claims, unless specifically defined in the claims where the term “means” is. The term “means” should not be construed to imply a relationship, but only in the claims, so-called “means plus function” only if specifically defined in a claim. Should be interpreted to mean a relationship. Similarly, these terms appearing in this application may be referred to as “step plus” in a method or process claim, unless the term “step” is specifically defined in the claims. function) ”relationship should not be construed as meaning, but only in the claims there is a so-called“ step ”in the claims only if the term“ step ”is specifically defined in the claims. plus function) ”relationship should be construed.

  Effect of fucoidan on synoviocyte and smooth muscle cell proliferation in vitro

  Proliferation was measured using a dimethylthiazole diphenyltetrazolium bromide salt (MTT) proliferation / cytotoxicity assay.

On the first day, 1500-2000 smooth muscle cells (A7r5 rat fetal thoracic aorta) or synoviocytes (HIG.82 rabbit) per well were placed in a 96-well plate. At that time, cells were not put in the wells in the first row of the plate and were blanked. The plate was set in a 37 ° C. CO ₂ incubator. The next day, various concentrations of fucoidan were added. At that time, fucoidan was not added to the blank column in the first row and the second column (untreated column) as a control. Cells were exposed for 48 hours. At the end of the exposure treatment, 50 μl of dimethylthiazole diphenyltetrazolium bromide salt (MTT) dissolved in a solvent was added and incubated at 37 ° C. for 4 hours. The medium was aspirated and 200 μl of dimethyl sulfoxide (DMSO) was added. The plate was shaken for 30 minutes and the absorbance was measured at 562 nm. Using a calibration curve of absorbance (optical density) of a sample with a known number of cells, the number of cells is determined from the measured value of absorbance, and the cell viability is determined as% proliferation (this value is% based on the control cell) Expressed in

  As shown in FIG. 1, fucoidan caused concentration-dependent cell growth inhibition after 48 hours exposure to both synoviocytes and smooth muscle cells. The inhibitory concentrations (IC50) showing 50% (inhibitory) action on proliferation (of synoviocytes and smooth muscle cells) were 15 μM and 6 μM, respectively.

  Effect of fucoidan on phorbol ester myristate (PMA) -induced neutrophil chemiluminescence

  In this experiment, freshly prepared human neutrophils were incubated with 0.5% w / v fucoidan and the cells were stimulated with PMA. This stimulation (or activation) of the cells produced a superoxide anion that could be measured by light emission (chemiluminescence). Inhibition of neutrophil function was determined by inhibition of chemiluminescence. In this experiment, a Hanks buffered saline solution (HBBS) having a pH of 7.4 was always used. All chemicals were purchased from Sigma Chemical Co (St. Louis, MO) unless otherwise specified. All experiments were performed at 37 ° C. Neutrophils were prepared from freshly collected human citrated whole blood. Briefly, 400 ml of blood was mixed with 80 ml of 4% dextran T500 (Pharmacia LKB, Biotechnology AB Uppsala, Sweden) in HBSS and allowed to stand for 1 hour. Plasma was continuously collected and added to 5 ml Ficoll Park (Pharmacia) in a 15 ml polypropylene tube (Corning, NY). After centrifugation at 500 × g for 30 minutes, the neutrophil pellet is washed with a 20 second hypotonic shock to remove red blood cells. Neutrophils were resuspended in HBSS, ice-cooled and used for experiments within 3 hours. Neutrophil viability and purity were always above 90%.

  Cells were incubated with various concentrations of fucoidan for 15 minutes at 37 ° C. before adding PMA.

The chemiluminescence test was performed at a cell concentration of 5 × 10 ⁶ cells / ml in HBBS containing 0.5 μM PMA. To each tube, 10 μL of luminol dissolved in 25% DMSO HBSS was added to a final concentration of 1 mM. Each sample was mixed to initiate neutrophil activation. Chemiluminescence was measured at 37 ° C. using an LKB luminometer (model 1250). Note that shaking was performed immediately before the measurement. The control tube contained cells, fucoidan and luminol.

  As shown in FIG. 2, fucoidan significantly inhibited PMA-induced neutrophil activation. Data are for three separate PMA-neutrophil culture samples. These data demonstrate the anti-inflammatory effect of fucoidan.

  Effect of fucoidan on IL-1-induced collagenase gene and stromelysin gene expression in chondrocytes

In this assay, the levels of RNA of two metalloproteinases, collagenase and stromelysin, are measured. Overexpression of these genes would secrete these two enzymes from articular chondrocytes and represent part of the pathophysiology of rheumatoid arthritis. Agents that inhibit the overexpression of collagenase and stromelysin are promising as anti-arthritic agents. If the agent also significantly inhibits proteoglycan gene expression, this potential as an anti-arthritic agent may be reduced. Proteoglycan gene expression is part of the normal physiology of chondrocytes. A primary culture of chondrocytes was freshly isolated from calf cartilage. Cells are transferred to a 100 × 20 mm culture dish (at 2.5 × 10 ⁶ / ml) and incubated overnight at 37 ° C. in Ham's F12 medium containing 5% fetal bovine serum (FBS). The cells were then starved overnight in serum free medium. Cells were then pretreated for 6 hours with camptothecin at concentrations of 10 ⁻⁶ M, 10 ⁻⁷ M and 10 ⁻⁸ M. IL-1 (20 ng / ml) was then added to each plate and the plate was further incubated for 18 hours. Total RNA was isolated by the acidified guanidine isothiocyanate method and subjected to electrophoresis on a denaturing gel. The denatured RNA samples (15 μg) were each subjected to gel electrophoresis using 1% denaturing gel, transferred to nylon membrane, ³² P-labeled collagenase cDNA probe, ³² P-labeled stromelysin cDNA probe, ³² P-labeled proteoglycan cDNA probe, respectively. , And ³² P-labeled glyceraldehyde phosphate dehydrogenase (PAGDH) cDNA probe. The PAGDH level was used as an internal standard to ensure approximately equal loading. The X-ray film experimental results were scanned and analyzed using HP ScanJet.

  As shown in FIG. 3, fucoidan completely inhibited collagenase and stromelysin expression at a concentration of 0.5% w / v without excessive inhibition of proteoglycan expression. Even at a concentration of 0.1% w / v, collagenase and stromelysin expression was strongly inhibited, but there was no inhibitory effect on proteoglycan expression. These data demonstrate the anti-inflammatory effect of fucoidan.

  Effect of fucoidan on angiogenesis in chorioallantoic membrane of chick embryo (CAM assay)

  Chicken fertilized eggs obtained from a local hatchery were placed in an incubator equipped with an automatic rotating device at 37 ° C. for 3.5 days, after which (partly) shell removal or window formation was performed.

  Several sterilized paraffin papers were placed on a window formed in the air chamber and used to prevent egg content contamination and dehydration (drying). This paraffin paper is 4 cm × 4 cm in size, but was sprayed with 70% ethanol, dried in a laminar flow draft and sterilized. Three days later, the eggs were manually rotated in an incubator so that their sharp tip was on top for 5-10 minutes and the egg contents were peeled off from the inner membrane. The entire eggshell was wiped with 70% ethanol and Kimwipe to clean and sterilize the egg surface. In a laminar flow draft, the egg was held with its blunt end facing up and an opening was formed at the blunt end of the egg by carefully cracking the shell with the tip of the tweezers. The shell residue was carefully removed with tweezers to form an opening at the blunt end. The diameter of this circular opening is about 2 to 3 cm, and the intima is not damaged. Immediately after forming an opening in the shell, gently soften the inner shell membrane (containing the egg contents), taking care not to damage the chorioallantoic membrane (CAM) (containing the yolk and the growing chick embryo) Break and remove with tweezers.

The opening was then covered with a piece of sterile parafilm (paraffin paper). At that time, the parafilm was gently stretched to cover the opening. The eggs were placed in an egg rack of an incubator (37 ° C.) and positioned so as not to roll. After 6 days, the eggs were removed from the incubator one at a time (with the blunt end facing up) and the parafilm covering the window was removed to allow direct access to the CAM. CAM has its origin in the hindgut of the embryo. Preparation of fucoidan-introduced poly (ε-caprolactone) (PCL) pellets was performed by melting PCL at 60 ° C., physically mixing fucoidan into the PCL, and cooling to room temperature to cure the pellets. Fucoidan pellets were placed on the growing capillary bed of the CAM. And the egg content was again sealed with the parafilm sheet | seat, and it returned to the 37 degreeC incubator. Two more days later, CAM vasculature analysis results were recorded (48 hours after application of the drug to the CAM capillary bed). The effect of the drug on CAM was evaluated using an avascular scale. In this method, the action and effect of the drug are evaluated as 0, 1, 2, and 3. Avascular grade values are shown below:
0 No anti-angiogenic activity 1 Microvascular reduction 2 Formation of small avascular region corresponding to drug pellet size (diameter 2 mm) 3 Formation of avascular region 4-5 mm in diameter

  As shown in Table 1, fucoidan strongly inhibited angiogenesis in CAM. Fucoidan inhibited CAM angiogenesis either partially (4 eggs) or completely (2 eggs) even at concentrations as low as 2% w / w in PCL.

Table 1: Anti-angiogenic activity of fucoidan. The numbers in each column indicate the number of eggs (CAM) that did not show any inhibition of angiogenesis, partially shown, or showed maximum.

Table 1

  These data demonstrate the anti-angiogenic activity of fucoidan and fucoidan's polymeric sustained release dosage form is an effective way to release pharmaceutically effective concentrations of drug without undue toxicity It is shown that.

  Encapsulation of fucoidan with ethylene vinyl acetate film and polycaprolactone paste (of fucoidan).

  5 mg of fucoidan (Sigma) and 45 mg of ethylene vinyl acetate (EVA, molecular weight of about 50 k, polysciences) were dissolved / suspended in 1 ml of dichloromethane. 200 μl of the solution was pipetted onto a 1 cm diameter Teflon disk and dried overnight (solvent evaporation) to form a thin stretch film. This film had a mass of about 10 mg and a thickness of about 100 μm.

  The drug release rate from the film was measured by putting 5 mg of the film fragment into a 20 ml lidded glass tube containing 10 ml of phosphate buffered saline (PBS) at pH 7.4. The lid of the glass tube was closed and set on an orbital shaker at 37 ° C. At predetermined time intervals, the glass tube was removed and the amount of drug released was analyzed by absorbance spectroscopy. From the fucoidan release curve (FIG. 4), it was found that the drug (fucoidan) was released rapidly at first but then continued to be released slowly. This dosage form of fucoidan represents a biocompatible, biodegradable injectable pharmaceutical dosage form that releases the drug in a sustained release manner.

  PCL paste: Fucoidan was mixed with polycaprolactone (PCL, Birmingham Polymers, molecular weight 54K) at a concentration of 10% w / w while being crushed with a spatula at 60 ° C. The mixture was then pipetted into a 1 ml plastic syringe and cooled. This dosage form could be injected by an 18 gauge needle at 56 ° C.

  To measure drug release from the PCL paste, a 10 mg aliquot of the molten paste was injected (injected) into the bottom of a 15 ml glass tube, cooled and set. Each glass tube was mixed by repeatedly adding 15 ml of PBS, closing the lid, and turning it upside down in an oven at 37 ° C. At each predetermined elapsed time, the glass tube was removed and the amount of drug released was analyzed by absorption spectroscopy. The release curve of fucoidan is shown in FIG. It was found that fucoidan is released rapidly at first, but then continues to be released slowly. This dosage form of fucoidan represents a biocompatible, biodegradable injectable pharmaceutical dosage form that releases the drug in a sustained release manner.

  Fucoidan-introduced membrane for treating surgical adhesions in rats

  A surgical adhesion model in the rat cecal wall was used to examine the effect of fucoidan on surgical adhesions. In this model, 16 rats were divided into two groups of 8 each. After the surgical trauma, one group of rats was immediately treated with cross-linked hyaluronic acid (HA) film containing fucoidan, and the other group of rats was not treated (control group).

Materials and methods . Medical grade sodium hyaluronate was obtained from Lifecore Scientific. All solvents were HPLC grade and were obtained from Fisher. Plastic petri dishes were obtained from Fisher Scientific. Ethyl-3- (dimethylamino) carbodiimide (EDAC) and fucoidan were obtained from Sigma (St. Louis, MO).

Film creation . To make a fucoidan-introduced film, an aqueous solution of 0.6% w / v fucoidan, 0.4% w / v sodium hyaluronate and 0.15% w / v glycerol was prepared. To make a control film (no fucoidan), an aqueous solution of 0.4% w / v sodium hyaluronate and 0.15% w / v glycerol was prepared. The fucoidan-introduced film and the control film were formed by first putting 4 g of each of these aqueous solutions with a pipette into separate 2.5 cm diameter plastic petri dishes and drying at 60 ° C. for 24 hours. The crosslinker EDAC was included (at final concentration) 4 mM. Next, each dried film was carefully peeled from the Petri dish using a scalpel.

Sterilization . The films were each wrapped between 5 cm × 5 cm weighing paper (Fisher Scientific), put in a plastic bag and heat sealed. The film was then finally sterilized by gamma radiation from a cobalt 60 source. The intensity of the radiation was 2.5 Mrad, and the radiation irradiation was performed while ice-cooling the sealed tube.

Animal experiment . Surgical trauma was formed as follows. Sixteen mature SD rats weighing 225-350 g were obtained from Charles River Laboratories, Wilmington, Massachusetts. Only rats that appeared to be very normal (i.e. cleanly with a jacket, bright and clear eyes and active behavior) were used in the experiment. Rats were randomly assigned to one of two groups, weighed, and anesthetized with a single injection of ketamine hydrochloride (6 mg / kg) into the thigh muscle. The abdomen was shaved and disinfected with alcohol. A 4 cm incision was made in the abdomen from the tail of about 2 cm to the white line, and the muscle was treated with forceps. The cecum was scraped 4 times with a mechanical scraping device on the ventral and back surfaces. This mechanical scraping device allows the surgeon to freely create a scraping in a desired manner in a specific area. The adhesion to the cecum was evaluated and graded according to the following evaluation criteria.
0 = No adhesion 1 = Thin (very weak) adhesion with easily recognizable (adhesion) surface 2 = Weak adhesion with freely separable (adhesion) surface 3 = Difficult to separate (adhesion) surface Medium adhesion 4 = Strong adhesion with non-separable (adhesion) surface (Stage 1 adhesion means the weakest level of recognizable adhesion (thin film with recognizable (adhesion) surface) adhesion))

  After rubbing the cecum, the first group of rats received no treatment. A second group of rats received the fucoidan-HA film treatment described above. The cecum was wrapped in this film. The incision was then closed using 3.0 Dexon thread. Seven days after surgery, the rats were euthanized and evaluated for the presence of stage 2 (or higher) post-surgical adhesions. Stage 2 adhesions are defined as weak adhesions with a freely detachable (adhesion) surface.

・盲腸は凡そ半分しか（フコイダン導入）膜で被覆しなかった。
・解剖の際異常は認められなかった（盲腸にも正中線切開部にも、残留物、腹水、異常な回復の徴候の何れも認められなかった）。 Result .

Table 2

• The cecum was only half covered (fucoidan introduced) with a membrane.
No abnormalities were observed during dissection (no residue, ascites, or signs of abnormal recovery were observed in the cecum or midline incision).

  This result shows that the average rate of adhesion formation is reduced and the percentage of rats with no adhesion formation is increased in rats treated with fucoidan, so that fucoidan-introduced (containing) films effectively inhibit adhesion formation. It has been proved that. If the cecum is completely covered with a fucoidan-introducing film, adhesion formation may be more effectively inhibited.

  While specific embodiments have been described herein, this is for illustrative purposes only and various modifications can be made without departing from the spirit and teachings of the present invention. Accordingly, the systems and methods of the present invention include such modifications and permutations / combinations of the subject matter defined herein, and are not limited except as by the appended claims.

The graph which shows the cell growth inhibition by the fucoidan with respect to a synovial cell and a smooth muscle cell after 48-hour exposure. The graph which shows inhibition of myristic acid phorbol ester (PMA) -induced neutrophil activation by fucoidan. Photograph showing fucoidan inhibition of collagenase and stromelysin expression at a concentration of 0.5% w / v without excessive inhibition of proteoglycan expression. The graph which shows the discharge | release of fucoidan from an ethylene vinyl acetate film. The graph which shows the discharge | release of fucoidan from a polycaprolactone paste.

Claims (70)

A method of treating adhesions in an animal, comprising administering a therapeutically effective amount of fucan to a disease site that may have adhesions. The method according to claim 1, wherein the fucan is fucoidan. The method according to claim 1, wherein the disease site is a surgical site, and the fucan is delivered directly to the disease site as a composition. The method according to any one of claims 1 to 3, wherein the fucan is administered from the polymer dosage form in a sustained manner and substantially continuously to the disease site. The method of claim 4, wherein the polymer dosage form comprises a film, patch, paste, microsphere, implant, gel, spray or liquid. 6. The fucan is administered as a pharmaceutical composition comprising at least one of a cream, a paste, an injectable excipient, and a polymer. The method described. The method according to any one of claims 1 to 6, wherein the fucan is administered as a pharmaceutical composition comprising the fucan and at least one therapeutically effective amount of another agent. 8. The method of claim 7, wherein the other agent comprises at least one of paclitaxel, doxorubicin, camptothecin, and etoposide. The other drug comprises at least one of mitoxantrone, methotrexate, menadione, plumbazine, juglone, β-lapachone cyclosporine, sulfasalazine, steroid, rapamycin, retinoid, docetaxel and colchicine. 8. The method according to 7. 8. The method of claim 7, wherein the other agent comprises at least one of an antisense oligonucleotide, a ribozyme, and an oligonucleotide RNA inhibitor. The therapeutically effective amount of fucan is delivered as part of a composition, wherein the fucan is included in the composition at about 20 to about 80% w / v (concentration). The method according to any one of 10 above. The therapeutically effective amount of fucan is delivered as part of a composition, the fucan being included in the composition at about 80 to about 100% w / v (concentration). The method according to any one of 10 above. The therapeutically effective amount of fucan is delivered as part of a composition, the fucan being included in the composition at about 5 to about 50% w / v (concentration). The method according to any one of 10 above. 14. The method according to any one of claims 1 to 13, wherein the composition further comprises at least one pharmaceutically acceptable excipient. 15. The method of claim 14, wherein the pharmaceutically acceptable excipient is selected from the group consisting of pluronic, cellulose, alginate, acrylate, hyaluronic acid, polyethylene glycol and chitosan. The method according to any one of claims 1 to 15, wherein the fucan is orally administered. The method according to any one of claims 1 to 16, wherein the fucan is directly administered to the disease site. The method according to any one of claims 1 to 17, wherein the fucan is administered to the disease site by injection. 19. The fucan is administered intraocularly, subcutaneously, intraperitoneally, intramuscularly, intraarticularly, intralesionally, intravaginally, rectally, or topically. The method according to claim 1. The method according to claim 1, wherein the animal is a human. A method for treating arthritis in an animal, comprising administering a therapeutically effective amount of fucan to a disease site containing arthritis. 24. The method of claim 23, wherein the arthritis is rheumatoid arthritis and the fucan is fucoidan. 23. The method according to claim 21 or 22, wherein the fucan is administered in a sustained manner from a polymer dosage form and substantially continuously to the disease site. 24. The method of claim 23, wherein the polymer dosage form comprises a film, patch, paste, microsphere, implant, gel, spray or liquid. The therapeutically effective amount of fucan is delivered as part of a composition, wherein the fucan is included in the composition at a concentration of about 0.1% to about 35% w / w. Item 25. The method according to any one of Items 21 to 24. 22. The therapeutically effective amount of fucan is delivered as part of a composition, wherein the fucan is included in the composition at a concentration of about 20% to about 80% w / v. The method according to any one of ˜25. 22. The therapeutically effective amount of fucan is delivered as part of a composition, wherein the fucan is included in the composition at about 5% to about 50% w / v (concentration). The method according to any one of -26. 28. The fucan is administered as a pharmaceutical composition comprising a form comprising at least one of a cream, a paste, an injectable excipient and a polymer. The method described. 29. A method according to any one of claims 21 to 28, wherein the composition further comprises at least one pharmaceutically acceptable excipient. 30. The method of claim 29, wherein the pharmaceutically acceptable excipient is selected from the group consisting of pluronic, cellulose, alginate, acrylate, hyaluronic acid, polyethylene glycol and chitosan. 31. The method of any one of claims 21-30, wherein the fucan is administered as a pharmaceutical composition comprising the fucan and at least one therapeutically effective amount of another agent. 32. The method of claim 31, wherein the other agent comprises at least one of paclitaxel, doxorubicin, camptothecin, and etoposide. The other drug comprises at least one of mitoxantrone, methotrexate, menadione, plumbazine, juglone, β-lapachone cyclosporine, sulfasalazine, steroid, rapamycin, retinoid, docetaxel and colchicine. 31. The method according to 31. 32. The method of claim 31, wherein the other agent comprises at least one of an antisense oligonucleotide, a ribozyme, and an oligonucleotide RNA inhibitor. The method according to any one of claims 21 to 34, wherein the fucan is administered orally. The method according to any one of claims 21 to 35, wherein the fucan is administered directly to the disease site. The method according to any one of claims 21 to 36, wherein the fucan is administered to the disease site by injection. 38. The fucan is administered intraocularly, subcutaneously, intraperitoneally, intramuscularly, intraarticularly, intralesionally, intravaginally, rectally, or topically. The method according to claim 1. The method according to any one of claims 21 to 38, wherein the animal is a human. A method for treating psoriasis, comprising administering a therapeutically effective amount of fucan to a disease site containing psoriasis. 41. The method of claim 40, wherein the fucan is fucoidan. 43. The method of claim 41 or 42, wherein the fucan is administered in a sustained manner from a polymer dosage form and substantially continuously to the disease site. 43. A method according to any one of claims 40 to 42, wherein the polymer dosage form is comprised of a film, patch, paste, microsphere, implant, gel, spray or liquid. 44. The fucan is administered as a pharmaceutical composition comprising a form comprising at least one of a cream, a paste, an injectable excipient, and a polymer. The method described. 45. The method of any one of claims 40 to 44, wherein the fucan is administered as a pharmaceutical composition comprising the fucan and at least one therapeutically effective amount of another agent. 46. The method of claim 45, wherein the other agent comprises at least one of paclitaxel, doxorubicin, camptothecin, and etoposide. The other drug comprises at least one of mitoxantrone, methotrexate, menadione, plumbazine, juglone, β-lapachone cyclosporine, sulfasalazine, steroid, rapamycin, retinoid, docetaxel and colchicine. 45. The method according to 45. 46. The method of claim 45, wherein the other agent comprises at least one of an antisense oligonucleotide, a ribozyme, and an oligonucleotide RNA inhibitor. The therapeutically effective amount of fucan is delivered as part of a composition, the fucan being included in the composition at a concentration of about 0.1% to about 35% w / w. Item 49. The method according to any one of Items 40 to 48. 41. The therapeutically effective amount of fucan is delivered as part of a composition, wherein the fucan is included in the composition at about 20% to about 80% w / v (concentration). 50. A method according to any one of -49. 41. The therapeutically effective amount of fucan is delivered as part of a composition, wherein the fucan is included in the composition at about 5% to about 50% w / v (concentration). The method according to any one of -50. 52. The method of any one of claims 40 to 51, wherein the composition further comprises at least one pharmaceutically acceptable excipient. 53. The method of claim 52, wherein the pharmaceutically acceptable excipient is selected from the group consisting of pluronic, cellulose, alginate, acrylate, hyaluronic acid, polyethylene glycol and chitosan. The method according to any one of claims 40 to 53, wherein the fucan is administered by a patch (patch). 55. The method according to any one of claims 40 to 54, wherein the fucan is administered directly to the disease site. The method according to any one of claims 40 to 55, wherein the fucan is administered locally. 57. The method according to any one of claims 40 to 56, wherein the animal is a human. A fucan polymeric dosage form comprising a therapeutically effective amount of fucan and at least one pharmaceutically acceptable excipient selected from the group consisting of pluronic, cellulose, alginate, acrylate, hyaluronic acid, polyethylene glycol and chitosan. A pharmaceutical composition comprising: 59. The composition of claim 58, wherein the polymeric dosage form is comprised of a film, patch, paste, microsphere, implant, gel, spray or liquid. 59. The composition of claim 58, wherein the fucan is fucoidan. Comprising a therapeutically effective amount of fucan and a therapeutically effective amount of at least one other agent, the other agent comprising at least one of an antisense oligonucleotide, a ribozyme and an oligonucleotide RNA inhibitor, A pharmaceutical composition. 62. The composition of claim 61, wherein the fucan is fucoidan.   63. A composition according to any one of claims 58 to 62 for use in the manufacture of a medicament for the treatment of adhesions. 64. The composition of claim 63, wherein the adhesion is an adhesion resulting from surgery.   63. A composition according to any one of claims 58 to 62 for use in the manufacture of a medicament for the treatment of arthritis.   63. A composition according to any one of claims 58 to 62 for use in the manufacture of a medicament for the treatment of psoriasis. Symptoms associated with at least one of adhesions, arthritis, and psoriasis in a human patient, comprising mixing a therapeutically effective amount of fucoidan and a pharmaceutically acceptable excipient or buffer. A method for producing a relaxable drug. The manufacturing method according to claim 67, wherein the symptom is associated with adhesion caused by surgery. 68. The method of claim 67, wherein the symptom is associated with arthritis. 68. The method of claim 67, wherein the symptom is associated with psoriasis.

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